心肌淀粉样变人血清对培养心肌细胞基因表达谱的影响  

Effect of human serum derived from cardiac amyloidosis on gene expression profiles in cultured cardiomyocytes

作　　者：许菲[1] 程仙[1] 王秀芝[1] 周芳[1] 张海峰[1] 周艳丽[1] 李新立[1] 陈相健[1] 徐东杰[1] XU Fei;CHENG Xian;WANG Xiuzhi;ZHOU Fang;ZHANG Haifeng;ZHOU Yanli;LI Xinli;CHEN Xiangjian;XU Dongjie(Department of Cardiology, First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029 ,China)

机构地区：[1]南京医科大学第一附属医院心脏内科,南京210029

出　　处：《临床心血管病杂志》2018年第6期613-619,共7页Journal of Clinical Cardiology

基　　金：卫生厅开放课题(No:XK05200903)

摘　　要：目的:利用基因表达谱芯片技术筛选正常人血清和心肌淀粉样变患者血清干预培养心肌细胞的差异表达基因,以探讨淀粉样变心肌病发病的可能机制。方法:通过将分离的血清稀释10倍后干预培养SD乳鼠心肌细胞建立模型,采用CCK-8检测各组细胞活性抑制程度,基因表达谱芯片技术筛选两组大鼠差异表达基因,采用实时定量PCR验证相关基因表达量。结果:CCK-8显示与正常control组相比,阴性对照组细胞存活率略有下降,但无统计学意义。与control组相比,实验组细胞存活率明显下降至约70%(P<0.05)。基因芯片分析显示,患者血清实验组及正常血清对照组两组相比,差异表达的基因有869条,其中上调基因710条,下调基因159条。实验组中胶原形成相关基因Col1a1、Col3a1、Col5a2、Col6a1、Col6a2、Col6a3,促纤维形成相关基因Lama1、Itga4、Vwf、Mmp14的表达均明显上调(P<0.05);影响血管重建相关基因Angptl4、Asb4、Cited1表达下调(P<0.05)。经RT-PCR验证,结果一致。结论:心肌淀粉样变患者血清干预影响培养心肌细胞的活性,促进胶原纤维形成并影响血管重建。淀粉样变心肌病的病理机制涉及多个基因,其中基质纤维化、血管重建受阻对淀粉样变心肌病的发生发展起着至关重要的作用,其具体机制还有待进一步研究。Objective：To explore the possible mechanism of cardiac amyloidosis.Method：The primary cultured cardiomyocytes from newborn SD rats were exposed with serum from normal person or patient with cardiac amyloidosis with 10 times dilution.The cytotoxicity of each group was detected by CCK-8 experiment.The differential genes expressions were screened by gene chip and the related genes were validated by Quantitative real-time-PCR.Result：CCK-8 showed that the cell viability was slightly decreased in the normal control group,compared the negative control group.Compared with control group,the survival rate of the experimental group decreased significantly to about 70%（P〈0.05）.Gene chip analysis showed that there were 869 differentially expressed genes between the serum test group and the normal serum control group,including 710 up-regulated genes and 159 down-regulated genes.The expression of collagen-forming genes Col1a1,Col3a1,Col5a2,Col6a1,Col6a2,Col6a3,and the genes related to the fibrogenesis-related genes Lama1,Itga4,Vwf and Mmp14 were significantly up-regulated in the experimental group（P〈0.05）.The expression of Angptl4,Asb4 and Cited1 were down-regulated（P〈0.05）.Conclusion：Serum intervention affects the activity of cultured cardiomyocytes in patients with myocardial amyloidosis.It promotes the alteration of genes related to collagen formation and vascular remodeling.

关 键 词：基因芯片 淀粉样变 纤维化 血管重建 

分 类 号：R542.2[医药卫生—心血管疾病]